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- Title
Cell cycle synchronization of canine ear fibroblasts for somatic cell nuclear transfer.
- Authors
Ok Jae Koo; Mohammad Shamim Hossein; So Gun Hong; Jose A. Martinez-Conejero; Byeong Chun Lee
- Abstract
SummaryCycle synchronization of donor cells in the G0/G1stage is a crucial step for successful somatic cell nuclear transfer. In the present report, we evaluated the effects of contact inhibition, serum starvation and the reagents ? dimethyl sulphoxide (DMSO), roscovitine and cycloheximide (CHX) ? on synchronization of canine fibroblasts at the G0/G1stage. Ear fibroblast cells were collected from a beagle dog, placed into culture and used for analysis at passages three to eight. The population doubling time was 36.5 h. The proportion of G0/G1cells was significantly increased by contact inhibition (77.1%) as compared with cycling cells (70.1%); however, extending the duration of culture did not induce further synchronization. After 24 h of serum starvation, cells were effectively synchronized at G0/G1(77.1%). Although synchronization was further increased gradually after 24 h and even showed significant difference after 72 h (82.8%) of starvation, the proportion of dead cells also significantly increased after 24 h. The percentage of cells at the G0/G1phase was increased (as compared with controls) after 72 h treatment with DMSO (76.1%) and after 48 h treatment with CHX (73.0%) or roscovitine (72.5%). However, the rate of cell death was increased after 24 and 72 h of treatment with DMSO and CHX, respectively. Thus, we recommend the use of roscovitine for cell cycle synchronization of canine ear fibroblasts as a preparatory step for SCNT.
- Subjects
CELL cycle regulation; SYNCHRONIZATION; FIBROBLASTS; SOMATIC cells; TRANSPLANTATION of cell nuclei; LABORATORY dogs; DIMETHYL sulfoxide
- Publication
Zygote, 2009, Vol 17, Issue 1, p37
- ISSN
0967-1994
- Publication type
Article
- DOI
10.1017/S096719940800498X